Construction and functional evaluation of an engineered bacte-rial strain for high-efficiency expression of NucB nuclease
Objective To objective of this study was to construct an engineered bacterial strain capable of effi-ciently and stably expressing NucB nuclease.Methods The DH5α(DE3)strain of Escherichia coli was used as the host organism.Gene knockout was performed using genetic editing techniques to disrupt the essential gene lexA within the bacterial genome.Subsequently,a modified plasmid containing the lexA gene was designed and introduced to complement the knockout,resulting in the engineered strain DH5α(DE3)ΔlexA.A plasmid carrying the target gene nucB was then transformed into the modified strain for fermentation cultivation and production of NucB nuclease,followed by functional characterization.Results A comparative analysis between the genetically modified strain DH5α(DE3)ΔlexA and the wild-type Escherichia coli revealed that the modi-fied strain exhibited highly stable plasmids,significantly increased protein expression levels,and the ability to produce NucB nuclease during fermentation without the need for antibiotic supplementation.Conclusion In summary,we successfully constructed an engineered strain by using genetic editing methods,DH5α(DE3)ΔlexA,which enables antibiotic-free,efficient,and stable expression of NucB nuclease.
gene knockoutantibiotic-freeefficient and stableNucB nuclease
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维普
